Abstract Since its introduction, in May 2015, the mosquito-borne Zika virus (ZIKV) has rapidly spread to more than 40 countries throughout the South Pacific and the Americas. The association between infection with ZIKV while pregnant and the occurrence of microcephaly in the fetus has raised a significant public health concern. To date, there is no effective antiviral treatment or vaccine to combat this rapidly escalating pandemic. The primary focus of this R21 application is to develop a novel Zika virus (ZIKV) capsid-capsid interaction assay and employ it to screen and identify chemical inhibitors of ZIKV replication. The formation of the ZIKV capsid protein dimer is a prerequisite to executing multiple functions in the virus lifecycle. Previous studies on other ZIKV-related C proteins have provided ample evidence that a relatively subtle perturbation in the efficiency and timing of C protein dimerization can disrupt proper viral assembly/disassembly and block virus infectivity. A recently described Dengue virus capsid assembly inhibitor supports the feasibility of our approach in exploring the ZIKV capsid as a target for anti-ZIKV drug discovery. Successful completion of this project will allow us develop a novel high-throughput assay for identifying ZIKV capsid assembly/disassembly inhibitors. The compounds identified through this work can be further studied to develop viable anti-ZIKV drug candidates that could be used for treatment of ZIKV infection in humans.